Automated vertical wheel bioreactor integrated with process analytics for T-cell manufacturing

  • Cytotherapy. 2025 Aug 31:S1465-3249(25)00813-8. doi: 10.1016/j.jcyt.2025.08.007.
Bryan Wang  1 Bharat Kanwar  2 Annie C Bowles-Welch  3 Walker Byrnes  4 Paloma Casteleiro Costa  5 Caroline Filan  5 Reginald Tran  6 Theresa Kotanchek  7 Francisco Robles  5 Krishnendu Roy  8 Stephen Balakirsky  9
Affiliations
  • 1. Marcus Center for Therapeutic Cell Characterization and Manufacturing, Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA; School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA; NSF Engineering Research Center (ERC) for cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA.
  • 2. Georgia Tech Research Institute, Georgia Institute of Technology, Atlanta, Georgia, USA; Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • 3. Marcus Center for Therapeutic Cell Characterization and Manufacturing, Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA; NSF Engineering Research Center (ERC) for cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA.
  • 4. Georgia Tech Research Institute, Georgia Institute of Technology, Atlanta, Georgia, USA; NSF Engineering Research Center (ERC) for cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA.
  • 5. Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.
  • 6. Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA; Department of Pediatrics, Division of Pediatric Hematology/Oncology, Emory University School of Medicine, Atlanta, Georgia, USA.
  • 7. Evolved Analytics LLC, Rancho Santa Fe, California, USA.
  • 8. Marcus Center for Therapeutic Cell Characterization and Manufacturing, Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA; NSF Engineering Research Center (ERC) for cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA. Electronic address: [email protected].
  • 9. Georgia Tech Research Institute, Georgia Institute of Technology, Atlanta, Georgia, USA; NSF Engineering Research Center (ERC) for cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA. Electronic address: [email protected].
Abstract

Background aims: Biomanufacturing of cell therapies involves highly complex and labor-intensive processes, where process parameters and biological variabilities can significantly influence product quality, reproducibility and therapeutic efficacy. Here, we developed a vertical wheel-based bioreactor platform with automated controls and in-line process analytical technologies (PAT) to demonstrate successful closed-system T cell biomanufacturing.

Methods: By identifying the critical process parameters (CPP), a process development strategy was optimized for expanding primary human unmodified and chimeric antigen receptor (CAR) T cells using multiple activation systems, including degradable microscaffolds.

Results: Spent media analysis combined with symbolic regression identified CPPs, which were validated through small-scale experiments and large-scale expansions in the bioreactor platform. Closed-loop automation with analytics such as real-time imaging also was integrated into the bioreactor platform for continuous monitoring and process control.

Conclusions: This integrated bioreactor platform provides a proof-of-concept design for multiplexed PAT integration, process optimization and feedback-controlled intelligent automation to enable discovery, monitoring and control of critical quality attributes and critical process parameters for cell therapy manufacturing.

Keywords
T cells; automation; bioreactor design; cell therapy manufacturing; process analytical technology; process development.
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